Product Info Summary
SKU: | PB9128 |
---|---|
Size: | 100 μg/vial |
Reactive Species: | Human, Mouse, Rat |
Host: | Rabbit |
Application: | Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-APE1/APEX1 Antibody Picoband™
SKU/Catalog Number
PB9128
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-APE1/APEX1 Antibody Picoband™ catalog # PB9128. Tested in Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.
Storage & Handling
Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
Cite This Product
Anti-APE1/APEX1 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9128)
Host
Rabbit
Contents
Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human APE1 recombinant protein (Position: P2-L318). Human APE1 shares 94% and 93% amino acid (aa) sequences identity with mouse and rat APE1, respectively.
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Cross-reactivity
No cross-reactivity with other proteins
Reactive Species
PB9128 is reactive to APEX1 in Human, Mouse, Rat
Applications
PB9128 is guaranteed for Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee
Observed Molecular Weight
39 kDa
Calculated molecular weight
35.555kDa
Background of APE
APEX1, also called apurinic endonuclease (APE), is a DNA repair enzyme having apurinic/apyrimidinic (AP) endonuclease, 3-prime, 5-prime-exonuclease, DNA 3-prime repair diesterase, and DNA 3-prime-phosphatase activities. The human APEX1 gene consists of 5 exons spanning 2.64 kb and exists as a single copy in the haploid genome. Using in situ hybridization, the APEX1 gene is mapped to 14q11.2-q12. The predicted APEX1 protein, which contained probable nuclear transport signals, was identified as a member of a family of DNA repair enzymes found in lower organisms. The abundance of the large form of APEX1 was increased in leiomyoma extracts relative to myometrial tissue extracts, and the large form was dominant in cell lines derived from leiomyosarcomas. The exonuclease activity of nuclear APEX1 can remove the anti-HIV nucleoside analogs AZT and D4T from the 3-prime terminus of a nick more efficiently than can cytosolic exonucleases.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
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Assay dilution & Images
Reconsitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.1-0.5μg/ml, Human, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, Mouse, Rat, By Heat
Immunocytochemistry/Immunofluorescence, 2 μg/ml, Human
Immunofluorescence, 2μg/ml, Human
Flow Cytometry, 1-3μg/1x106 cells, Human
Validation Images & Assay Conditions
Click image to see more details
Figure 1. Western blot analysis of APEX1 using anti-APEX1 antibody (PB9128).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours.
Lane 1: Recombinant Human APEX1 Protein 0.5ng.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-APEX1 antigen affinity purified polyclonal antibody (Catalog # PB9128) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for APEX1 at approximately 45KD. The expected band size for APEX1 is at 45KD.
Click image to see more details
Figure 2. Western blot analysis of APEX1 using anti-APEX1 antibody (PB9128).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: NRK Whole Cell Lysate
Lane 2: HELA Whole Cell Lysate
Lane 3: PC-12 Whole Cell Lysate
Lane 4: RH35 Whole Cell Lysate
Lane 5: HEPA Whole Cell Lysate
Lane 6: MCF Whole Cell Lysate
Lane 7: A549 Whole Cell Lysate
Lane 8: Human Placenta Tissue Lysate
Lane 9: A431 Whole Cell Lysate
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-APEX1 antigen affinity purified polyclonal antibody (Catalog # PB9128) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for APEX1 at approximately 39KD. The expected band size for APEX1 is at 39KD.
Click image to see more details
Figure 3. IHC analysis of APEX1 using anti-APEX1 antibody (PB9128).
APEX1 was detected in paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-APEX1 Antibody (PB9128) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 4. IHC analysis of APEX1 using anti-APEX1 antibody (PB9128).
APEX1 was detected in paraffin-embedded section of mouse brain tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-APEX1 Antibody (PB9128) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 5. IHC analysis of APEX1 using anti-APEX1 antibody (PB9128).
APEX1 was detected in paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-APEX1 Antibody (PB9128) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 6. IF analysis of APEX1 using anti-APEX1 antibody (PB9128).
APEX1 was detected in immunocytochemical section of U20S cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti-APEX1 Antibody (PB9128) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Click image to see more details
Figure 7. Flow Cytometry analysis of U937 cells using anti-APEX1 antibody (PB9128).
Overlay histogram showing U937 cells stained with PB9128 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-APEX1 Antibody (PB9128,1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Click image to see more details
Figure 8. IF analysis of APEX1 using anti-APEX1 antibody (PB9128).
APEX1 was detected in paraffin-embedded section of human colon cancer tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/mL rabbit anti-APEX1 Antibody (PB9128) overnight at 4°C.DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Protein Target Info & Infographic
Gene/Protein Information For APEX1 (Source: Uniprot.org, NCBI)
Gene Name
APEX1
Full Name
DNA-(apurinic or apyrimidinic site) lyase
Weight
35.555kDa
Superfamily
DNA repair enzymes AP/ExoA family
Alternative Names
AP endonuclease 1; APE; APE1; APE-1; APEAPEX nuclease (multifunctional DNA repair enzyme); APENAP endonuclease class I; APEX deoxyribonuclease (apurinic or apyrimidinic); APEX nuclease (multifunctional DNA repair enzyme) 1; APEX nuclease; APEX1; Apurinic-apyrimidinic endonuclease 1; APXAP lyase; DNA-(apurinic or apyrimidinic site) lyase; EC 3.1; EC 4.2.99.18; HAP1apurinic/apyrimidinic (abasic) endonuclease; multifunctional DNA repair enzyme; protein REF-1; redox factor 1; Redox factor-1; REF1 apurinic/apyrimidinic exonuclease; REF-1 APEX1 APE, APE1, APEN, APEX, APX, HAP1, REF1 apurinic/apyrimidinic endodeoxyribonuclease 1 DNA-(apurinic or apyrimidinic site) endonuclease|AP endonuclease class I|AP lyase|APEX nuclease (multifunctional DNA repair enzyme) 1|DNA-(apurinic or apyrimidinic site) lyase|apurinic-apyrimidinic endonuclease 1|apurinic/apyrimidinic (abasic) endonuclease|deoxyribonuclease (apurinic or apyrimidinic)|protein REF-1|redox factor-1
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on APEX1, check out the APEX1 Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for APEX1: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-APE1/APEX1 Antibody Picoband™ (PB9128)
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Customer Q&As
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1 Customer Q&As for Anti-APE1/APEX1 Antibody Picoband™
Question
We are currently using anti-APE1/APEX1 antibody PB9128 for rat tissue, and we are happy with the IHC-P results. The species of reactivity given in the datasheet says human, mouse, rat. Is it possible that the antibody can work on pig tissues as well?
Verified Customer
Verified customer
Asked: 2017-12-20
Answer
The anti-APE1/APEX1 antibody (PB9128) has not been validated for cross reactivity specifically with pig tissues, but there is a good chance of cross reactivity. We have an innovator award program that if you test this antibody and show it works in pig you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2017-12-20